Expression and nucleotide sequence of the Clostridium acetobutylicum β-galactosidase gene cloned in Escherichia coli

Article


Hancock Kerrie R., Rockman, Eva, Young, Carolyn A., Pearce, Lindsay, Maddox, Ian S. and Scott, Barry. 1991. "Expression and nucleotide sequence of the Clostridium acetobutylicum β-galactosidase gene cloned in Escherichia coli." Journal of Bacteriology. 173 (10), pp. 3084-3095. https://doi.org/10.1128/jb.173.10.3084-3095.1991
Article Title

Expression and nucleotide sequence of the Clostridium acetobutylicum β-galactosidase gene cloned in Escherichia coli

ERA Journal ID2483
Article CategoryArticle
AuthorsHancock Kerrie R., Rockman, Eva, Young, Carolyn A., Pearce, Lindsay, Maddox, Ian S. and Scott, Barry
Journal TitleJournal of Bacteriology
Journal Citation173 (10), pp. 3084-3095
Number of Pages11
Year1991
PublisherAmerican Society for Microbiology
Place of PublicationUnited States
ISSN0021-9193
1067-8832
1098-5530
Digital Object Identifier (DOI)https://doi.org/10.1128/jb.173.10.3084-3095.1991
Web Address (URL)https://journals.asm.org/doi/10.1128/jb.173.10.3084-3095.1991
Abstract

A gene library for Clostridium acetobutylicum NCIB 2951 was constructed in the broad-host-range cosmid pLAFR1, and cosmids containing the β-galactosidase gene were isolated by direct selection for enzyme activity on X-Gal (5-bromo-4-chloro-3-indoly-β-D-galactoside) plates after conjugal transfer of the library to a lac deletion derivative of Escherichia coli. Analysis of various pSUP202 subclones of the lac cosmids on X-Gal plates localized the β-galactosidase gene to a 5.1-kb EcoRI fragment. Expression of the Clostridium β-galactosidase gene in E. coli was not subject to glucose repression. By using transposon Tn5 mutagenesis, two gene loci, cbgA (locus I) and cbgR (locus II), were identified as necessary for β-galactosidase expression in E. coli. DNA sequence analysis of the entire 5.1-kb fragment identified open reading frames of 2,691 and 303 bp, corresponding to locus I and locus II, respectively, and in addition a third truncated open reading frame of 825 bp. The predicted gene product of locus I, CbgA (molecular size, 105 kDa), showed extensive amino acid sequence homology with E. coli LacZ, E. coli EbgA, and Klebsiella pneumoniae LacZ and was in agreement with the size of a polypeptide synthesized in maxicells containing the cloned 5.1-kb fragment. The predicted gene product of locus II, CbgR (molecular size, 11 kDa) shares no significant homology with any other sequence in the current DNA and protein sequence data bases, but Tn5 insertions in this gene prevent the synthesis of CbgA. Complementation experiments indicate that the gene product of cbgR is required in cis with cbgA for expression of β-galactosidase in E. coli.

ANZSRC Field of Research 2020310505. Gene expression (incl. microarray and other genome-wide approaches)
PubMed ID1850729
Byline AffiliationsMassey University, New Zealand
Permalink -

https://research.usq.edu.au/item/w3v61/expression-and-nucleotide-sequence-of-the-clostridium-acetobutylicum-galactosidase-gene-cloned-in-escherichia-coli

  • 4
    total views
  • 0
    total downloads
  • 1
    views this month
  • 0
    downloads this month

Export as

Related outputs

Addition of Activated Carbon into a Cattle Diet to Mitigate GHG Emissions and Improve Production
Al-Azzawi, Mohammed, Bowtell, Les, Hancock, Kerry and Preston, Sarah. 2021. "Addition of Activated Carbon into a Cattle Diet to Mitigate GHG Emissions and Improve Production." Sustainability. 13 (15), pp. 1-17. https://doi.org/10.3390/su13158254
Progress in the pathobiology and genetic epidemiology of Campylobacter hepaticus, the aetiological agent of Spotty Liver Disease
Kotiw, M., Courtice, J., Mahdi, L., Wei, C., Ahmad, T., Hancock, K. and Groves, P.. 2019. "Progress in the pathobiology and genetic epidemiology of Campylobacter hepaticus, the aetiological agent of Spotty Liver Disease." 2019 Australasian Veterinary Poultry Association (AVPA) Annual Scientific Meeting. Sydney, Australia 20 - 21 Feb 2019
Elevation of Condensed Tannins in the Leaves of Ta-MYB14-1 White Clover (Trifolium repens L.) Outcrossed with High Anthocyanin Lines
Roldan, Marissa B., Cousins, Greig, Fraser, Karl, Hancock, Kerry R., Collette, Vern, Demmer, Jerome, Woodfield, Derek R., Caradus, John R., Jones, Chris and Voisey, Christine R.. 2020. "Elevation of Condensed Tannins in the Leaves of Ta-MYB14-1 White Clover (Trifolium repens L.) Outcrossed with High Anthocyanin Lines." Journal of Agricultural and Food Chemistry. 68 (10), pp. 2927-2939. https://doi.org/10.1021/acs.jafc.9b01185
Breaking free: The genomics of allopolyploidy-facilitated niche expansion in white clover
Griffiths, Andrew G., Moraga, Roger, Tausen, Marni, Gupta, Vikas, Bilton, Timothy P., Campbell, Matthew A., Ashby, Rachael, Nagy, Istvan, Khan, Anar, Larking, Anna, Anderson, Craig, Franzmayr, Benjamin, Hancock Kerry, Scott, Alicia, Ellison,Nick W., Cox, Murray P., Asp, Torben, Mailund, Thomas, Schierup, Mikkel H. and Andersen, Stig Uggerhøj. 2019. "Breaking free: The genomics of allopolyploidy-facilitated niche expansion in white clover." The Plant Cell. 31 (7), pp. 1466-1487. https://doi.org/10.1105/tpc.18.00606
Characterization of Proanthocyanidins from Seeds of Perennial Ryegrass (Lolium perenne L.) and Tall Fescue (Festuca arundinacea) by Liquid Chromatography-Mass Spectrometry
Fraser, Karl, Collette, Vern and Hancock Kerry R.. 2016. "Characterization of Proanthocyanidins from Seeds of Perennial Ryegrass (Lolium perenne L.) and Tall Fescue (Festuca arundinacea) by Liquid Chromatography-Mass Spectrometry." Journal of Agricultural and Food Chemistry. 64 (35), pp. 6676-6684. https://doi.org/10.1021/acs.jafc.6b02563
Progress towards developing bloat-safe legumes for the farming industry
Hancock, Kerry, Collette, Vern, Chapman, Elisabeth, Hanson, Katherine, Temple, Stephen, Moraga, Roger and Caradus, John. 2014. "Progress towards developing bloat-safe legumes for the farming industry." Crop and Pasture Science. 65 (11), pp. 1107-1113. https://doi.org/10.1071/CP13308
An integrated genetic linkage map for white clover (Trifolium repens L.) with alignment to Medicago
Griffiths, Andrew G, Barrett, Brent A, Simon, Deborah, Khan, Anar K, Bickerstaff, Paul, Anderson, Craig B, Franzmayr, Benjamin K, Hancock Kerry R. and Jones, Chris S. 2013. "An integrated genetic linkage map for white clover (Trifolium repens L.) with alignment to Medicago." BMC Genomics. 14 (1), pp. 1-17. https://doi.org/10.1186/1471-2164-14-388
Expression of the R2R3-MYB transcription factor TaMYB14 from Trifolium arvense Activates proanthocyanidin biosynthesis in the legumes Trifolium repens and Medicago sativa
Hancock, Kerry R., Collette, Vern, Fraser, Karl, Greig, Margaret, Xue, Hong, Richardson, Kim, Jones, Chris and Rasmussen, Susanne. 2012. "Expression of the R2R3-MYB transcription factor TaMYB14 from Trifolium arvense Activates proanthocyanidin biosynthesis in the legumes Trifolium repens and Medicago sativa." Plant Physiology. 159 (3), pp. 1204-1220. https://doi.org/10.1104/pp.112.195420
Fructan 2000
Biggs, David R. and Hancock, Kerrie R.. 2001. "Fructan 2000." Trends in Plant Science. 6 (1), pp. 8-9. https://doi.org/10.1016/S1360-1385(00)01796-9
Expression of a sulfur-rich maize seed storage protein, δ-zein, in white clover (Trifolium repens) to improve forage quality
Sharma, Shashi B., Hancock Kerrie R., Ealing, Paul M. and White, Derek W.R.. 1998. "Expression of a sulfur-rich maize seed storage protein, δ-zein, in white clover (Trifolium repens) to improve forage quality." Molecular Breeding: new strategies in plant improvement. 4 (5), pp. 435-448. https://doi.org/10.1023/A:1009656002068
In vitro digestion of bacterial and plant fructans and effects on ammonia accumulation in cow and sheep rumen fluids
Biggs, David R. and Hancock Kerrie R.. 1998. "In vitro digestion of bacterial and plant fructans and effects on ammonia accumulation in cow and sheep rumen fluids." The Journal of General and Applied Microbiology. 44 (2), pp. 167-171. https://doi.org/10.2323/jgam.44.167
pPE1000: A versatile vector for the expression of epitope-tagged foreign proteins in transgenic plants
Hancock Kerrie R., Phillips, Lorelle D., White, Derek W.R. and Ealing, Paul M.. 1997. "pPE1000: A versatile vector for the expression of epitope-tagged foreign proteins in transgenic plants." BioTechniques: the journal of laboratory technology for bioresearch. 22 (5), pp. 861-863. https://doi.org/10.2144/97225bm16
Identification of sulphur-rich proteins which resist rumen degradation and are hydrolysed rapidly by intestinal proteases
Hancock Kerrie R., Ealing, Paul M. and White, Derek W.R.. 1994. "Identification of sulphur-rich proteins which resist rumen degradation and are hydrolysed rapidly by intestinal proteases." The British Journal of Nutrition: an international journal of nutritional science. 72 (6), pp. 855-863. https://doi.org/10.1079/BJN19940090
Expression of the pea albumin 1 gene in transgenic white clover and tobacco
Ealing, Paul M., Hancock Kerrie R. and White, Derek D. W.. 1994. "Expression of the pea albumin 1 gene in transgenic white clover and tobacco." Transgenic Research. 3 (6), pp. 344-354. https://doi.org/10.1007/BF01976766